Non-invasive surface-stripping for epifluorescence small animal imaging

Area of Science:

• Biomedical Imaging
• Optical Imaging
• Medical Physics

Background:

• Non-invasive near-infrared fluorescence (NIRF) imaging is valuable for studying animal disease models, particularly brain diseases.
• Superficial fluorescence from scalp and blood vessels can hinder the detection of deeper tissue signals in NIRF imaging.
• Targeting deeper tissues requires methods to overcome signal attenuation and background interference.

Purpose of the Study:

• To develop and validate an effective method for reducing superficial fluorescence interference in NIRF imaging.
• To enhance the contrast and detectability of fluorochromes in deeper tissue layers.
• To improve the diagnostic capabilities of NIRF imaging for deep-seated pathologies.

Main Methods:

• Utilizing excitation light at two distinct wavelengths to generate images with differential depth sensitivity.
• Implementing an adapted subtraction algorithm to process the generated images.
• Testing the technique in tissue-simulating phantoms and murine models of stroke.

Main Results:

• Significant enhancement in image contrast was achieved.
• Improved detectability of fluorochromes located in deep tissue layers was demonstrated.
• The method effectively reduced the impact of superficial fluorescence.

Conclusions:

• The developed two-wavelength excitation and subtraction algorithm method significantly improves deep tissue NIRF imaging.
• This technique offers a promising approach for non-invasive visualization of deep pathologies in preclinical research.
• Enhanced NIRF imaging can advance the study of neurological diseases and other deep-seated conditions.